Since the O.lurida RNA I isolated on 20150507 showed residual gDNA via qPCR, I treated 1.5μg of RNA from each sample using the Turbo DNA-free Kit (Ambion/Life Technologies), following the “rigorous” protocol.
Briefly:
50μL reactions were carried out in 0.5mL tubes
added 1μL of DNase to each tube
incubated 30mins @ 37C
added additional 1μL of DNased
incubated 30mins @ 37C
added 0.2 vols (10.2μL) of DNase Inactivation Reagent
incubated and mixed for 2mins @ RT
transferred 50μL of supe to sterile 1.5mL snap cap tubes
spec’d on Roberts Lab NanoDrop1000
Samples were stored @ -80C in Shellfish RNA Box #5 and Box #6.
DNase reaction calcs: 20150514_Jake_Oly_1hr_HS_DNase_calcs
Results:
Google Spreadsheet: 20150514_DNased_RNA_Jake_Oly_1hr_HS_ODs
(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_ODs.JPG)
(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_01.JPG)
(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_02.JPG)
All samples look pretty good except for HT1 8 (RNA concentration is ridiculously high!) and NT1 8 (RNA concentration is way below expected). Will check for residual gDNA via qPCR.